Anti-sirp alpha antibodies

Abstract

The present invention relates to anti-SIRPα antibodies, as well as use of these antibodies in the treatment of diseases such as cancer and infectious disease.

1 - 30 . (canceled) 
     
     
         31 . An antibody antibody having the following characteristics:
 binds human SIRPαV1 protein having the sequence of SEQ ID NO: 34 with an EC 50 <1 nM;   does not cross-react with SIRPβ1 protein having the sequence of SEQ ID NO: 38; and   exhibits a T20 “humanness” score of at least 79.   
     
     
         32 . An antibody according to  claim 31 , wherein the antibody binds to a cell expressing human SIRPαV1 protein with an EC 50 <10 nM;
 binds to a cell expressing human SIRPαV2 protein with an EC 50 <10 nM; 
 exhibits at least a 100-fold higher EC 50  for SIRPαV1(P74A) having the sequence of SEQ ID NO: 62 as compared to the EC 50  for human SIRPαV1 protein; and 
 exhibits at least a 100-fold higher EC 50  for human SIRPβ1 protein as compared to the EC 50  for human SIRPαV1 protein. 
 
     
     
         33 . An antibody according to  claim 31 , wherein the antibody inhibits binding between human SIRPα and CD47 with an IC 50 <2.5 nM. 
     
     
         34 . An antibody according to  claim 32 , wherein the antibody inhibits binding between human SIRPα and CD47 with an IC 50 <2.5 nM. 
     
     
         35 . A composition comprising:
 an antibody or antigen binding fragment of  claim 31 ; and   a pharmaceutically acceptable carrier or diluent,   wherein the composition optionally comprises a second antibody or antigen binding fragment thereof that induces ADCC and/or ADCP, wherein said antibody or antigen binding fragment of  claim 31  enhances the antibody-mediated destruction of cells by the second antibody.   
     
     
         36 . A composition comprising:
 an antibody or antigen binding fragment of  claim 31 ; and   a pharmaceutically acceptable carrier or diluent,   
     
     
         37 . A composition according to  claim 36 , further comprising a second antibody or antigen binding fragment thereof that induces ADCC and/or ADCP. 
     
     
         38 . An antibody or antigen binding fragment thereof that binds to the same epitope of human SIRPα as an antibody comprising one of the following combinations of heavy chain sequence/light chain sequence:
 SEQ ID NO: 78/SEQ ID NO: 90, 
 SEQ ID NO: 78/SEQ ID NO: 92, 
 SEQ ID NO: 78/SEQ ID NO: 94, 
 SEQ ID NO: 78/SEQ ID NO: 96, 
 SEQ ID NO: 78/SEQ ID NO: 98, 
 SEQ ID NO: 78/SEQ ID NO: 100, 
 SEQ ID NO: 80/SEQ ID NO: 90, 
 SEQ ID NO: 80/SEQ ID NO: 92, 
 SEQ ID NO: 80/SEQ ID NO: 94, 
 SEQ ID NO: 80/SEQ ID NO: 96, 
 SEQ ID NO: 80/SEQ ID NO: 98, 
 SEQ ID NO: 80/SEQ ID NO: 100, 
 SEQ ID NO: 82/SEQ ID NO: 90, 
 SEQ ID NO: 82/SEQ ID NO: 92, 
 SEQ ID NO: 82/SEQ ID NO: 94, 
 SEQ ID NO: 82/SEQ ID NO: 96, 
 SEQ ID NO: 82/SEQ ID NO: 98, 
 SEQ ID NO: 82/SEQ ID NO: 100, 
 SEQ ID NO: 84/SEQ ID NO: 90, 
 SEQ ID NO: 84/SEQ ID NO: 92, 
 SEQ ID NO: 84/SEQ ID NO: 94, 
 SEQ ID NO: 84/SEQ ID NO: 96, 
 SEQ ID NO: 84/SEQ ID NO: 98, 
 SEQ ID NO: 84/SEQ ID NO: 100, 
 SEQ ID NO: 86/SEQ ID NO: 90, 
 SEQ ID NO: 86/SEQ ID NO: 92, 
 SEQ ID NO: 86/SEQ ID NO: 94, 
 SEQ ID NO: 86/SEQ ID NO: 96, 
 SEQ ID NO: 86/SEQ ID NO: 98, 
 SEQ ID NO: 86/SEQ ID NO: 100, 
 SEQ ID NO: 88/SEQ ID NO: 90, 
 SEQ ID NO: 88/SEQ ID NO: 92, 
 SEQ ID NO: 88/SEQ ID NO: 94, 
 SEQ ID NO: 88/SEQ ID NO: 96, 
 SEQ ID NO: 88/SEQ ID NO: 98, 
 SEQ ID NO: 88/SEQ ID NO: 100, 
 SEQ ID NO: 10/SEQ ID NO: 20, 
 SEQ ID NO: 10/SEQ ID NO: 22, 
 SEQ ID NO: 10/SEQ ID NO: 24, 
 SEQ ID NO: 10/SEQ ID NO: 26, 
 SEQ ID NO: 10/SEQ ID NO: 28, 
 SEQ ID NO: 12/SEQ ID NO: 20, 
 SEQ ID NO: 12/SEQ ID NO: 22, 
 SEQ ID NO: 12/SEQ ID NO: 24, 
 SEQ ID NO: 12/SEQ ID NO: 26, 
 SEQ ID NO: 12/SEQ ID NO: 28, 
 SEQ ID NO: 14/SEQ ID NO: 20, 
 SEQ ID NO: 14/SEQ ID NO: 22, 
 SEQ ID NO: 14/SEQ ID NO: 24, 
 SEQ ID NO: 14/SEQ ID NO: 26, 
 SEQ ID NO: 14/SEQ ID NO: 28, 
 SEQ ID NO: 16/SEQ ID NO: 20, 
 SEQ ID NO: 16/SEQ ID NO: 22, 
 SEQ ID NO: 16/SEQ ID NO: 24, 
 SEQ ID NO: 16/SEQ ID NO: 26, 
 SEQ ID NO: 16/SEQ ID NO: 28, 
 SEQ ID NO: 18/SEQ ID NO: 20, 
 SEQ ID NO: 18/SEQ ID NO: 22, 
 SEQ ID NO: 18/SEQ ID NO: 24, 
 SEQ ID NO: 18/SEQ ID NO: 26, 
 SEQ ID NO: 18/SEQ ID NO: 28. 
 
     
     
         39 . An isolated nucleic acid encoding an antibody or antigen binding fragment thereof that binds to human SIRPα, wherein the antibody or antigen binding fragment comprises:
 a. a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:69 or an amino acid sequence differing from SEQ ID NO: 1 by 1, 2, or 3 conservative substitutions, 
 b. a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:70 or an amino acid sequence differing from SEQ ID NO: 2 by 1, 2, or 3 conservative substitutions, 
 c. a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:71 or an amino acid sequence differing from SEQ ID NO: 3 by 1, 2, or 3 conservative substitutions, 
 d. a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:72 or an amino acid sequence differing from SEQ ID NO: 4 by 1, 2, or 3 conservative substitutions, 
 e. a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:73 or an amino acid sequence differing from SEQ ID NO: 5 by 1, 2, or 3 conservative substitutions, and 
 f. a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:74 or an amino acid sequence differing from SEQ ID NO: 6 by 1, 2, or 3 conservative substitutions. 
 
       or wherein the antibody or antigen binding fragment comprises:
 g. a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:1 or an amino acid sequence differing from SEQ ID NO: 1 by 1, 2, or 3 conservative substitutions, 
 h. a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:2 or an amino acid sequence differing from SEQ ID NO: 2 by 1, 2, or 3 conservative substitutions, 
 i. a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:3 or an amino acid sequence differing from SEQ ID NO: 3 by 1, 2, or 3 conservative substitutions, 
 j. a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:4 or an amino acid sequence differing from SEQ ID NO: 4 by 1, 2, or 3 conservative substitutions, 
 k. a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:5 or an amino acid sequence differing from SEQ ID NO: 5 by 1, 2, or 3 conservative substitutions, and 
 l. a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:6 or an amino acid sequence differing from SEQ ID NO: 6 by 1, 2, or 3 conservative substitutions. 
 
     
     
         40 . An expression vector comprising the isolated nucleic acid of  claim 39 . 
     
     
         41 . A host cell comprising expression vector of  claim 40 . 
     
     
         42 . A method of producing an antibody or antigen binding fragment comprising:
 culturing a host cell comprising a polynucleotide of  claim 39  under conditions favorable to expression of the polynucleotide; and optionally, recovering the antibody or antigen binding fragment from the host cell and/or culture medium.   
     
     
         43 . A method for detecting the presence of a SIRPα peptide or a fragment thereof in a sample comprising contacting the sample with an antibody or fragment an antibody or antigen binding fragment thereof that binds to human SIRPα, wherein the antibody or antigen binding fragment comprises:
 a. a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:69 or an amino acid sequence differing from SEQ ID NO: 1 by 1, 2, or 3 conservative substitutions, 
 b. a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:70 or an amino acid sequence differing from SEQ ID NO: 2 by 1, 2, or 3 conservative substitutions, 
 c. a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:71 or an amino acid sequence differing from SEQ ID NO: 3 by 1, 2, or 3 conservative substitutions, 
 d. a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:72 or an amino acid sequence differing from SEQ ID NO: 4 by 1, 2, or 3 conservative substitutions, 
 e. a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:73 or an amino acid sequence differing from SEQ ID NO: 5 by 1, 2, or 3 conservative substitutions, and 
 f. a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:74 or an amino acid sequence differing from SEQ ID NO: 6 by 1, 2, or 3 conservative substitutions. 
 
       or wherein the antibody or antigen binding fragment comprises:
 g. a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:1 or an amino acid sequence differing from SEQ ID NO: 1 by 1, 2, or 3 conservative substitutions, 
 h. a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:2 or an amino acid sequence differing from SEQ ID NO: 2 by 1, 2, or 3 conservative substitutions, 
 i. a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:3 or an amino acid sequence differing from SEQ ID NO: 3 by 1, 2, or 3 conservative substitutions, 
 j. a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:4 or an amino acid sequence differing from SEQ ID NO: 4 by 1, 2, or 3 conservative substitutions, 
 k. a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:5 or an amino acid sequence differing from SEQ ID NO: 5 by 1, 2, or 3 conservative substitutions, and 
 l. a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:6 or an amino acid sequence differing from SEQ ID NO: 6 by 1, 2, or 3 conservative substitutions; and 
 
       detecting the presence of a complex between the antibody or fragment and the peptide; wherein detection of the complex indicates the presence of the SIRPα peptide. 
     
     
         44 . A method of treating cancer, an infection, or infectious disease in a human subject, comprising administering to the subject an effective amount of an antibody or antigen binding fragment of  claim 31 . 
     
     
         45 . A method of treating cancer, an infection, or infectious disease in a human subject, comprising administering to the subject an effective amount of an antibody or antigen binding fragment of  claim 32 .